I-and II-type crystals of L-A-glyceryl phosphoryl choline, and method for preparing same

ABSTRACT

The present invention relates to I- and II-type crystals of L-α-glyceryl phosphoryl choline, and to a method for preparing same. More particularly, the present invention relates to noble I- and II-type anhydride crystals of L-α-glyceryl phosphoryl choline, which have a higher purity than conventional liquid L-α-glyceryl phosphoryl choline, and one advantage of which is that formulations and dosages of pharmaceuticals are easily modified, and another advantage of which is that the hygroscopicity of the crystals are much lower than that of conventional polymorphic crystals, providing excellent stability during storage. The present invention also relates to a method for preparing the I- and II-type crystals of L-α-glyceryl phosphoryl choline. The I-type crystal of L-α-glyceryl phosphoryl choline is characterized by having an onset temperature of 147° C. and an absorption peak of 150° C. based on differential scanning calorimeter (DSC) analysis, and by having diffraction patterns where 2θ-diffraction angles are 9.8±0.2°, 12.0±0.2°, 14.3±0.2°, 15.8±0.2°, and 19.6±0.2° based on X-ray diffraction (XRD) analysis. The II-type crystal of L-α-glyceryl phosphoryl choline is characterized by having an onset temperature of 62° C. and an absorption peak of 66° C., and an onset temperature of 141° C. and an absorption peak of 145° C. based on DSC analysis, and having diffraction patterns where 2θ-diffraction angles are 10.3±0.2°, 12.2±0.2°, 13.4±0.2°, 14.8±0.2°, and 20.6±0.2° based on XRD analysis.

CROSS-REFERENCES TO RELATED APPLICATION

This application is a Divisional Application of U.S. patent applicationSer. No. 14/003,276 filed Sep. 5, 2013, which is a National Stageapplication of PCT/KR2012/001331 filed on Feb. 22, 2012, which claimspriority to Korean Patent Application Nos. 10-2011-0022392 filed on Mar.14, 2011 and 10-2011-0022393 filed on Mar. 14, 2011, the contents ofwhich are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to type I and type II crystals ofL-alpha-glyceryl phosphoryl choline and to a method for preparing same.More specifically, the present invention relates to preparation of newtype I and type II crystals of L-alpha-glyceryl phosphoryl choline whichhave more advantages than the conventional liquid L-alpha-glycerylphosphoryl choline: The purity is higher, the formulations and dosagesare more easily modified in the sense of manufacturing pharmacy, and thestorage stability is better due to a much lower hygroscopic propertythan conventional polymorphic crystals.

BACKGROUND TECHNOLOGY

L-alpha-glyceryl phosphoryl choline (GPC hereunder) expressed by thefollowing chemical formula I is a substance known to be used to improvebrain function and treat dementia.

The conventional method of preparing L-alpha-glyceryl phosphoryl cholineis largely divided into the pure synthetic method and the extractionmethod in which the choline is extracted from recithin, a by-product ofbean. For example, the methods introduced in such patents as Europeanpatent No. 486.100, Italian Patent No. 1,243,724, and Italian Patent No.1,247,496 are the pure synthetic method, and the methods introduced insuch patents as U.S. Pat. No. 5,250,719, British Patent No. 2,058,792,and European Patent No. 217,765 are the recithin extraction method. Theconventional GPC prepared with these methods is obtained at the liquidstate containing a considerable amount of water.

Meanwhile, the method for crystallization of the above-mentioned liquidL-alpha-glyceryl phosphoryl choline is also known. First of all, in J.Am. Chem. Soc. 70, 1394-1399 (1948), it was reported that thewater-containing GPC prepared with the pure synthetic method could besolidified from the alcohol solution, but the detailed crystallizationmethod and the crystalline structure were not mentioned.

In Korean Patent No. 262281 (registration date: Apr. 29, 2000), a newmethod of preparing GPC was introduced: First, GPC was synthesized bythe alcoholysis deacylation reaction in a reactor containing a basic ionexchange resin and the lipophilic impurities are removed by using anon-polar adsorption resin. Then, the above-mentioned GPC was dissolvedin methanol and n-butanol whose amount was 20 times the methanolsolution was added to the methanol solution. Finally, the total solutionwas concentrated in vacuum, cooled, and filtered to recover an anhydrousform of crystal. In this method, however, though only the formation ofvery hygroscopic fine crystals was reported, details on the crystallinestructure were not mentioned.

And, in Korean Patent No. 966,627 (registration date: Jun. 20, 2010), amethod of preparing GPC hydrochloride, as expressed by chemical formula2, by using a crystallization solvent composed of C1-C6 alcohols, C1-C6ketones and their mixtures, was introduced but again the crystallinestructure was not mentioned.

Like this, though efforts have been made to crystallize GPC and thepossibility of crystallization has been proposed, details on thecrystallization conditions or the crystalline structure has not beenmentioned. And the raw material of crystalline GPC available forindustrial applications has not produced yet.

DETAILED DESCRIPTION OF INVENTION Technical Problem

As observed above, all the raw materials of GPC which have been producedindustrially so far are at the liquid state and the GPC medicines undercurrent sale are limited to soft capsules made by using theabove-mentioned liquid raw material. In such soft capsules, however, theactive component is likely to dissolve the gelatin capsula as the timepasses, complex manufacturing facilities are required, and not only apreserving agent should be used to prevent microbial degeneration butalso they are inconvenient for patients to take.

Thus, while conducting research on powder-type GPC in order tomanufacture GPC tablets which may replace GPC soft capsules, the presentinventors confirmed that all the GPC crystals obtained with thecrystallization method known so far are polymorphic ones and that thereexist new Type I and Type II crystals which have totally differentcrystalline properties from the above-mentioned polymorphic crystal. Andthe present invention was completed by developing an effective method ofproducing industrially each of the above-mentioned Type I and Type IIcrystals on the basis of the use of seed crystal and the optimizedcrystallization conditions.

The objective of the present invention is to provide the Type I and TypeII crystals of L-alpha-glyceryl phosphoryl choline and the method ofpreparing same.

Technical Solution

According to the present invention, Type I crystal of L-alpha-glycerylphosphoryl choline is characteristic in that it has an onset temperatureof 147° C. and an absorption peak of 150° C. on the basis of thedifferential scanning calorimetry (DSC) analysis and it also hasdiffraction patterns where 2θ-diffraction angles are 9/8±0.2°,12.0±0.2°, 14.3±0.2°, 15.8±0.2°, and 19.6±0.2° on the basis of thepowder X-ray diffraction (XRD) analysis.

In addition, the manufacturing method of the above-mentioned Type Icrystal is characterized by; A) a step of decreasing the water contentto 6-12% by concentrating ordinary liquid L-alpha-glyceryl phosphorylcholine at 45-65° C.; B) a step of dissolving the concentratedL-alpha-glyceryl phosphoryl choline obtained from the step A) intoalcohol solution whose amount is 1 to 5 times that of theabove-mentioned concentrated choline and then cooling the entiresolution to 5-20° C.; and C) a step of feeding, as the seed crystal,0.1-0.5 mole % of L-alpha-glyceryl phosphoryl choline Type I crystalinto the alcohol solution of L-alpha-glyceryl phosphoryl cholineobtained from the step B) process, aging the mixed solution while it isstirred at 30-60 rpm for 2 to 5 hours, and then filtering the extractedcrystal.

According to the present invention, Type II crystal of L-alpha-glycerylphosphoryl choline is characteristic in that it has an onset temperatureof 62° C./an absorption peak of 66° C. and an onset temperature of 141°C./an absorption peak of 145° C. on the basis of the differentialscanning calorimetry (DSC) analysis and it also has diffraction patternswhere 2θ-diffraction angles are 10.3±0.2°, 12.2±0.2°, 13.4±0.2°,14.8±0.2°, and 20.6±0.2° on the basis of the powder X-ray diffraction(XRD) analysis.

In addition, the manufacturing method of the above-mentioned Type IIcrystal is characterized by; A) a step of decreasing the water contentto 6-12% by concentrating ordinary liquid L-alpha-glyceryl phosphorylcholine at 45-65° C.; B) a step of dissolving the concentratedL-alpha-glyceryl phosphoryl choline obtained from the step A) processinto alcohol solution whose amount is 1 to 5 times that of theabove-mentioned concentrated choline and then cooling the entiresolution to 5-20° C.; and C) a step of feeding, as the seed crystal,0.1-0.5 mole % of L-alpha-glyceryl phosphoryl choline Type II crystalinto the alcohol solution of L-alpha-glyceryl phosphoryl cholineobtained from the step B), aging the mixed solution without agitationfor 2 to 5 hours and then filtering the extracted crystal.

Advantageous Effects

According to the present invention, Type I and Type II crystals ofL-alpha-glyceryl phosphoryl choline are effective in producing a varietyof medicines with high drug compliance to patients since the purity ishigher and the formulations and dosages are easier in the sense ofmanufacturing pharmacy than those of the conventional liquid rawmaterial of L-alpha-glyceryl phosphoryl choline.

In addition, the above-mentioned crystals have advantages that thestorage stability is excellent and they are easy to handle during themedication process due to much lower hygroscopic property than theconventional polymorphic crystal.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 X-ray diffraction spectrum of GPC Type I crystal in the presentinvention

FIG. 2 X-ray diffraction spectrum of GPC Type II crystal in the presentinvention

FIG. 3 X-ray diffraction spectrum of conventional GPC polymorphiccrystal

FIG. 4 Differential scanning calorimetric spectrum of GPC Type I in thepresent invention

FIG. 5 Differential scanning calorimetric spectrum of GPC Type II in thepresent invention

FIG. 6 Differential scanning calorimetric spectrum of conventional GPCpolymorphic crystal

FIG. 7 Graphs for comparison in hygroscopic property between GPC Type I,GPC Type II and polymorphic crystal

FIG. 8 IR spectrum of GPC Type I, GPC Type II and polymorphic crystal

BEST MODES FOR CARRYING OUT THE INVENTION

Type I crystal of L-alpha-glyceryl phosphoryl choline of the presentinvention is characteristic in that it has, as an anhydride, an onsettemperature of 147° C. and an absorption peak of 150° C. in thedifferential scanning calorimetry (DSC) analysis and has diffractionpatterns wherein 2θ-diffraction angles are 9/8±0.2°, 12.0±0.2°,14.3±0.2°, 15.8±0.2°, and 19.6±0.2° in the powder X-ray diffraction(XRD) analysis.

The above-mentioned Type I crystal of L-alpha-glyceryl phosphorylcholine has the tetrahedral structure of the tetragonal system in whichthe size of crystal grain is uniform at about 95-115 μm and theconsolute temperature is about 150° C. (1 degree/minute). And since thehygroscopicity is low in comparison with the conventional polymorphiccrystal, though the raw material with a water content of 0.1% is leftunder a humidity condition of 30% for about 10 hours the water contentincreases only slightly to 0.2%.

In addition, Type II crystal of L-alpha-glyceryl phosphoryl choline is,as a mono-hydrate, characteristic in that it has an onset temperature of62° C./an absorption peak of 66° C. and an onset temperature of 141°C./an absorption peak of 145° C. on the basis of the differentialscanning calorimetry (DSC) analysis and it also has diffraction patternswhere 2θ-diffraction angles are 10.3±0.2°, 12.2±0.2°, 13.4±0.2°,14.8±0.2°, and 20.6±0.2° on the basis of the powder X-ray diffraction(XRD) analysis.

The above-mentioned Type II crystal of L-alpha-glyceryl phosphorylcholine has the polyhedral form of the hexagonal system in which thesize of crystal grain is uniform at out 200-300 μm and the consolutetemperature is about 66° C. (1 degree/minute). And since thehygroscopicity is low in comparison with the conventional polymorphiccrystal, though the raw material with a water content of 6.5% is leftunder a humidity condition of 30% for about 10 hours, the water contentremains almost unchanged at 6.5%.

For reference, the conventional polymorphic crystal is relatively veryhygroscopic. In case the raw material with a water content of 2.9% isleft under a humidity condition of 30%, as time passes, it absorbsmoisture from air to reach a water content of 4.2% after 10 hours. Ifthe hygroscopic property is large like this, special handling requiredsince the weight and physical property are likely to change during theraw material storage or the medication process.

The preparation method of the above-mentioned Type I and Type IIcrystals of L-alpha-glyceryl phosphoryl choline consists of thefollowing 3 steps:

A) Concentration Step of Liquid L-Alpha-Glyceryl Phosphoryl Choline

First of all, ordinary L-alpha-glyceryl phosphoryl choline isconcentrated to decrease the water content to 6-12%. At this time, ifthe above-mentioned concentration temperature is below 45° C. or above65° C., lamp black substances are created to cause difficulties duringthe purification process. And if the water content after concentrationis below 6%, the undesirable polymorphic crystal may be generated.Conversely, if it is above 12%, the yield of crystallization falls agreat deal, which is undesirable.

For reference, the moisture content for the raw material of ordinaryliquid L-alpha-glyceryl phosphoryl choline is about 15-18%. Such liquidraw material may be used without concentration. In this case, however,since the yield of crystallization is low, it is desirable toconcentrate it to a water content of 6-12%.

In case the liquid L-alpha-glyceryl phosphoryl choline with a watercontent of 15-18% is used as the starting material, the water contentdecreases to 6-12% if it is concentrated at 45-65° C. for 8-10 hours.

B) Dissolution Step of Concentrated L-Alpha-Glyceryl Phosphoryl Choline

Next, the L-alpha-glyceryl phosphoryl choline concentrated in theabove-mentioned A) step is fed and dissolved into an alcohol solution,the foreign materials are removed by filtering the mixed solution with amembrane filter, and then it is cooled to 5-20° C.

At this time, ethanol and isopropyl alcohol may be used as theabove-mentioned alcohol solution for crystallization and the alcoholamount used is 1 to 5 times the amount of the concentratedL-alpha-glyceryl phosphoryl choline, preferably 2 to 5 times. If theabove-mentioned alcohol solution amount used is out of theabove-mentioned range, the crystallization efficiency decreases, whichresults in reduction in the yield and purity.

In addition, if the cooling temperature of the above-mentioned alcoholsolution is below 5° C., the size of crystal grain does not grow enoughdue to a very fast crystal formation. Conversely, if it is above 20° C.,GPC remains in the residual water and alcohol after concentration, whichcauses the crystallization yield to decrease.

C) Crystallization Step

Type I and Type II crystals are separated by the final crystallizationstep. First, if 0.1-0.5 mol % of the Type I crystal of L-alpha-glycerylphosphoryl choline is added, as the seed crystal, to the alcoholsolution of L-alpha-glyceryl phosphoryl choline obtained from theabove-mentioned B) step, the solution is aged while stirred at 30-60 rpmfor 2 to 5 hours, and then the extracted crystal is filtered, Type Icrystal of anhydrous form is obtained.

According to the results of the present inventors, it may be possible toobtain Type I crystal without feeding the above-mentioned seed crystal.In this case, however, the above-mentioned alcohol solution should becooled below −5° C. and the size of crystal grain does not grow enough.In addition, care should be taken since Type II crystal is also formedtogether if agitation is not provided during the above-mentioned agingprocess. The first Type I crystal acquired by the present inventors wasprepared without using the seed crystal.

Meanwhile, if 0.1-0.5 mol % of the Type II crystal of L-alpha-glycerylphosphoryl choline is added, as the seed crystal, to the alcoholsolution of L-alpha-glyceryl phosphoryl choline obtained from theabove-mentioned B) step, the solution is aged without agitation for 2 to5 hours, and then the extracted crystal is filtered, Type II crystal ofmonohydrate form is obtained.

According to the results of the present inventors, it may be possible toobtain Type II crystal without feeding the above-mentioned seed crystal.In this case, however, the aging time takes more than 24 hours and thesize of crystal grain does not grow enough. In addition, care should betaken since Type I crystal is also extracted together if agitation isprovided during the above-mentioned aging process. The first Type IIcrystal acquired by the present inventors was prepared without using theseed crystal.

Like this, in the present invention, around the seed crystal during theaging process, the size of crystal grain grows to 95-115 μm in case ofType I crystal and to 200-300 μm in case of Type II crystal. If the sizeof crystal grain grows like this, the hygroscopic property relativelydecreases due to reduction in the total surface area of powder.

Hereunder, desirable examples are described in order to assist inunderstanding of the present invention. However, the right extent of thepresent invention is not limited due to the examples mentioned below.

EXAMPLE 1

The liquid L-alpha-glyceryl phosphoryl choline with a water content of18% (1.5 kg) was concentrated in vacuum at 50° C. for 9 hours todecrease the water content to about 7%. Here, 3 liter of ethanol was fedand agitation was provided, while the temperature was maintained at 50°C., for complete dissolution. And then impurities were removed by using1.0 μm membrane filter.

Next, after the above-mentioned L-alpha-glyceryl phosphoryl cholinesolution was cooled to 9° C. and 2 g of Type I crystal ofL-alpha-glyceryl phosphoryl choline was fed as the seed crystal, thesolution was aged for 2 hours with an agitation of 50 rpm. The crystalextracted this way was filtered and vacuum-dried to obtain 1.04 kg(yield: 85%) of Type I crystal of L-alpha-glyceryl phosphoryl cholinewith a water content of 0.1%.

EXAMPLE 2

The liquid L-alpha-glyceryl phosphoryl choline with a water content of18% (1.5 kg) was concentrated in vacuum at 50° C. to decrease the watercontent to about 9%. Here, 3 liter of ethanol was fed and agitation wasprovided, while the temperature was maintained at 50° C., for completedissolution. And then impurities were removed by using 1.0 μm membranefilter

Next, after the L-alpha-glyceryl phosphoryl choline solution was cooledto 10° C. and 2 g of Type II crystal of L-alpha-glyceryl phosphorylcholine was fed as the seed crystal, the solution was aged for 4 hourswithout agitation. The crystal extracted this way was filtered andvacuum-dried to obtain 1.02 kg (yield: 83%) of Type II crystal ofL-alpha-glyceryl phosphoryl choline with a water content of 6.5%.

COMPARATIVE EXAMPLE

The liquid L-alpha-glyceryl phosphoryl choline (30 g) with a watercontent of 18% was concentrated in vacuum at 50° C. and 60 g of ethanolwas added for azeotropic distillation to reduce the water content to 4%,which was confirmed. The solution was then cooled to 5° C. and theformed crystal was aged for 1 hour. Then, the extracted crystal wasfiltered and dried in vacuum to obtain 17 g (yield: 69%) of thepolymorphic crystal of L-alpha-glyceryl phosphoryl choline with a watercontent of 2.9%.

For reference, in this comparison Example, the crystallization method inJ. Am. Chem. Soc. 70, 1394-1399 (1948) and Korean Patent No. 262,281 wasimplemented.

[Crystal Analysis and Property Test]

A. Powder X-Ray Diffraction (XRD) Analysis

According to the results for the powder X-ray diffraction (XRD) analysison the GPC crystal obtained in the above-mentioned examples, the2θ-diffraction angle of Type I crystal showed peaks at 9/8±0.2°,12.0±0.2°, 14.3±0.2°, 15.8±0.2°, and 19.6±0.2° as shown in FIG. 1 andthat of Type II crystal at 10.3±0.2°, 12.2±0.2°, 13.4±0.2°, 14.8±0.2°,and 20.6±0.2° as shown in FIG. 2.

In addition, according to the results for the powder X-ray diffraction(XRD) analysis on the GPC polymorphic crystal obtained in theabove-mentioned comparison Example, the 2θ-diffraction angle showedpeaks at 9.8±0.2°, 12.0±0.20°, 13.4±0.2°, 14.3±0.2°, 14.8±0.2°,15.8±0.2°, and 19.9±0.2°.

The measurement conditions for the above-mentioned powder X-raydiffraction (XRD) spectrum are as follows:

1) Equipment: PANalytical, X'Pert-Pro/X-Ray Source: Cu

2) Tube Voltage: 40 kV/Tube Current: 30 mA

3) Emission Slot: ½°/Scattering Slot: ½°/Light Receiving Slot: 0.15 mm

4) Scanning Zone: 5 to 40° 2θ/Sampling Interval: 0.02°

5) Scanning Speed: 0.02°/second

B. Differential Scanning calorimetry (DSC) Analysis

According to the results for the differential scanning calorimetry (DSC)analysis on the GPC crystal obtained in the above-mentioned examples,Type I crystal showed an onset temperature of 147° C. and an absorptionpeak of 150° C. as shown in FIG. 3, and Type II crystal showed an onsettemperature of 62° C./an absorption peak of 66° C. and an onsettemperature of 141° C./an absorption peak of 145° C. as shown in FIG. 5.

According to the results for the differential scanning calorimetry (DSC)analysis on the GPC polymorphic crystal obtained in the above-mentionedcomparison, the polymorphic crystal showed an onset temperature of 60°C./an absorption peak of 60.4° C., an onset temperature of 62° C./anabsorption peak of 64° C., an onset temperature of 143° C./an absorptionpeak of 146° C., and an onset temperature of 147° C./an absorption peakof 149° C. as shown in FIG. 6.

The measurement conditions for the above-mentioned differential scanningcalorimetry (DSC) spectrum are as follows:

1) Equipment: Mettler DSC 1102-R081,X

2) Measurement Range: 50 to 200° C./Temperature Rising Interval: 1°C./min

3) N2 Speed: 50 ml/min

C. Microphotography

The results for the microphotography (magnification: ×100) of theabove-mentioned GPC crystals confirmed that Type I crystal has thetetrahedral form of the tetragonal system in which each crystal graingrows uniformly in a size of 95 to 115 μm, forming a smooth grainsurface, and Type II crystal has the polyhedral form of the hexagonalsystem in which each crystal grain grows uniformly in a size of 200 to300 μm, also forming a smooth gain surface.

However, it was confirmed that, in the polymorphic crystal obtained inthe above-mentioned comparison, the crystal grain with a size of 20 to25 μm agglomerates to make a lump with a size of 150 to 160 μm, formingrough bumps on the surface.

D. Hygroscopicity Test

FIG. 7 shows the graphs for the water content change with the passage oftime for GPC Type I crystal, GPC Type II crystal, and polymorphiccrystal which are left under a humidity condition of 30% for 10 hours.In the figure, X axis indicates the elapse time and Y axis the watercontent. The above-mentioned Type I crystal and Type II crystalmaintained the initial water contents as they are even after passage of10 hours, indicating a very low hygroscopicity.

In case of the polymorphic crystal, however, the initial water contentwas 2.9% but increased to 4.2% after 10 hours due to absorption moisturefrom air. In comparison with Type I and Type II crystals, the highhygroscopicity of the polymorphic crystal is considered to result fromrough bumps on the crystal surface.

E. Infrared (IR) Absorption Spectrum

Finally, FIG. 8 shows the measurement for each infrared (IR) spectrum ofGPC Type I crystal, GPC Type II crystal and the polymorphic crystal.While Type I and Type II crystals have their unique absorption spectra,the polymorphic crystal is found to possess all the characteristics ofType I and II crystals.

The measurement conditions for the above-mentioned infrared (IR)absorption spectrum are as follows:

1) Equipment: FT/IR-4100 (Jasco)

2) Measurement Range: 4000 to 650 cm-1

3) Resolution: 4.00 cm-1

4) Number of Scans: 36

The invention claimed is:
 1. Type II crystal of L-alpha-glycerylphosphoryl choline, wherein an onset temperature is 62° C. and anabsorption peak is 66° C., and an onset temperature is 141° C. and anabsorption peak is 145° C., respectively, in a differential scanningcalorimetry (DSC) analysis, wherein 2θ-diffraction angles of thediffraction pattern are 10.3±0.2°, 12.2±0.2°, 13.4±0.2°, 14.8±0.2°, and20.6±0.2° in an X-ray diffraction (XRD) analysis, and wherein the TypeII crystal of L-alpha-glyceryl phosphoryl choline, as a mono-hydrate,has a polyhedral structure of a hexagonal system.
 2. A method ofmanufacturing the Type II crystal of L-alpha-glyceryl phosphoryl cholineof claim 1, the method comprising: A) decreasing water content ofordinary L-alpha-glyceryl phosphoryl choline to 6-12% by concentratingthe ordinary L-alpha-glyceryl phosphoryl choline at 45-65° C.; B)dissolving the concentrated L-alpha-glyceryl phosphoryl choline obtainedin step A) in an alcohol solution, an amount of the alcohol solutionbeing 1 to 5 times of an amount the concentrated choline and thencooling the entire solution to 5-20° C.; and C) feeding, as a seedcrystal, 0.1 to 0.5 mol % of Type II of L-alpha-glyceryl phosphorylcholine into the alcohol solution of L-alpha-glyceryl phosphoryl cholineobtained in step B), aging the entire solution for 2-5 hours withoutagitation, and then filtering the extracted crystal.
 3. The methodaccording to claim 2, wherein the concentrated L-alpha-glycerylphosphoryl choline of step A) has the water content of 15 to 18%originally and is concentrated for 8 to 10 hours.
 4. The methodaccording to claim 2, wherein the concentrated L-alpha-glycerylphosphoryl choline of step B) is dissolved in ethanol or isopropanol, anamount of the ethanol or isopropanol being 2 to 5 times an amount of theconcentrated choline.